Fluorescence photobleaching

Petersen N. O. and Elson E. L. (1986) Measurements of Diffusion and Chemical Kinetics by Fluorescence Photobleaching Recovery and Fluorescence Correlation Spectroscopy Methods Enzymol. 130, 454-84. 

D. Axelrod, D. E. Koppel, J. Schlessmjer, E. Elson, and W. W. Webb, Mobility measurement by analysis of fluorescence photobleaching recovery kinetics, Biophys. J. 16, 1055-1069... 

To increase the speed of the TIRF-based kinetic techniques, the perturbation can be optical rather than chemical. If the evanescent wave intensity is briefly flashed brightly, then some of the fluorophores associated with the surface will be photobleached. Subsequent exchange with unbleached dissolved fluorophores in equilibrium with the surface will lead to a recovery of fluorescence, excited by a continuous but much attenuated evanescent wave. The time course of this recovery is a measure of the desorption kinetic rate k2. This technique1-115) is called TIR/FRAP (or TIR/FPR) in reference to fluorescence recovery after photobleaching (or fluorescence photobleaching recovery). 

R. M. Zimmermann, C. F. Schmidt, and H. E. Gaub, Absolute quantities and equilibrium kinetics of macromolecular adsorption measured by fluorescence photobleaching in total internal reflection, J. Colloid Interface Sci. 139, 268-280 (1990). 

T. P. Burghardt and D. Axelrod, Total internal reflection/fluorescence photobleaching recovery study of serum albumin adsorption dynamics, Biophys. J. 33, 455-468 (1981). 

M. M. Timbs and N. L. Thompson, Slow rotational mobilities of antibodies and lipids associated with substrate-supported phospholipid monolayers as measured by polarized fluorescence photobleaching recovery, Biophys. J. 58, 413-428 (1990). 

Chary, S.R. and Jain, R.K. (1989) Direct measurement of interstitial convection and diffusion of albumin in normal and neoplastic tissues by fluorescence photobleaching. Proc. Natl. Acad. Sci. USA, 86, 5385-5389. 

Kaufman, E.N. and Jain, R.K. (1992) Effect of bivalent interaction upon apparent antibody affinity experimental confirmation of theory using fluorescence photobleaching and implications for antibody binding assays. Cancer Res., 52,4157 1167. 

D. M. Soumpasis, Theorectical-analysis of fluorescence photobleaching recovery experiments, Biophys. J. 41, 95-97 (1983). 

C. Lagerholm, T. E. Starr, Z. N. Volovyk, and N. L. Thompson, Rebinding of IgE Fabs at haptenated planar membranes Measurement by total internal reflection with fluorescence photobleaching recovery, Biochemistry 39, 2042-2051 (2000). 

Yu and Russo 52a reported the fluorescence photobleaching recovery and dynamic light... 

Thompson NL, Steele BL. Total internal reflection with fluorescence correlation spectroscopy. Nat. Protoc. 2007 2 878-890. Sund SE, Axelrod D. Actin dynamics at the living cell submembrane imaged by total internal reflection fluorescence photobleach-ing. Biophys. J. 2000 79 1655-1669. 

A new technique for measuring equilibrium adsorption/desorption kinetics and surface diffusion of fluorescent-labelled solute molecules at surfaces was developed by Thompson et al.74). The technique combines total internal reflection fluorescence with either fluorescence photobleaching recovery or fluorescence correlation spectroscopy with lasers. For example, fluorescent labelled protein was studied in regard to the surface chemistry of blood 75). 

Robinson, D.J., de Bruijn, H.S., van der Veen, N., Stringer, M.R., Brown, S.B., and Star, W.M. (1998) Fluorescence photobleaching of ALA-induced protoporphyrin IX during photodynamic therapy of normal hairless mouse skin the effect of light dose and irradiance and the resulting biological effect, Photochem. Photobiol., 67 140-149. 

Figure 10. Mutual diffusion coefficients, of the two molecular populations of a fluorescent dye dispersed in poly(1-trimethyl-l-propyne) plotted as functions of reciprocal absolute temperature. The technique used was laser fluorescence photobleaching recovery.

Wey, C., and Cone, R. A., Lateral diffusion of rhodopsin in photoreceptor cells measured by fluorescence photobleaching and recovery. Biophys. J. 33, 225 (1981). 

Fig. 8. Interstitial velocity profiles. Representative regions in the microcirculation. Circles represent locations of fluorescence photobleaching experiments. The arrows inside the circles represent the direction of the interstitial fluid velocity at these locations. The nearby values show magnitudes of the velocity in fim/s. (a) An area where interstitial flow parallels blood flow in the vessels, (b) Interstitial flow is opposite prevailing blood flow, (c) Fluid is absorbed from the interstitium into a postcapillary venule. (From Chary and Jain, 1989, with permission.) The photobleaching technique has provided the first and to date the only measurements in the literature of interstitial convective velocities. We have now further improved this technique to permit measurements of binding parameters (Kaufman and Jain, 1990, 1991, 1992a, b) and of transport parameters in light-scattering media (Berk et al., 1993).

Berk, D.A., et al. Fluorescence photobleaching with spatial fourier analysis Measurement of diffusion in light-scattering media. Biophysical Journal, 1993, 65, 2428-2436. 

Barisas, B.G. and M.D. Leuther, Fluorescence photobleaching recovery measurement of protein absolute diffusion constants. Biophysical Chemistry, 1977,10, 221-229. 

W.R. Zipfel, W.W.W. Webb, In vivo Diffusion Measurements Using Multiphoton Excitation Fluorescence Photobleaching Recovery and Fluorescence Correlation Spectroscopy, in A. Periasamy (ed.), Methods in Cellular Imaging, Oxford University Press, 216 (2001)... 

Another exciting technique combines FRAP with total internal reflection fluorescence microscopy. This technique, called total internal reflection interference fringe fluorescence photobleaching... 

Hagen, G. M., Roess, D. A., and Barisas, B. G. (2006) Fluorescence photobleaching recovery using total internal reflection interference fringes. Anal. Biochem. 356, 30-35. 

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